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Applied and Environmental Microbiology, April 2004, p. 2013-2020, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2013-2020.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Protective Effect of Sucrose and Sodium Chloride for Lactococcus lactis during Sublethal and Lethal High-Pressure Treatments

Adriana Molina-Höppner,1 Wolfgang Doster,2 Rudi F. Vogel,1 and Michael G. Gänzle*

Lehrstuhl für Technische Mikrobiologie, Technische Universität München, D-85350 Freising,1 Lehrstuhl E13, Fakultät für Physik, Technische Universität München, 85748 Garching, Germany2

Received 16 June 2003/ Accepted 20 December 2003

The bactericidal effect of hydrostatic pressure is reduced when bacteria are suspended in media with high osmolarity. To elucidate mechanisms responsible for the baroprotective effect of ionic and nonionic solutes, Lactococcus lactis was treated with pressures ranging from 200 to 600 MPa in a low-osmolarity buffer or with buffer containing 0.5 M sucrose or 4 M NaCl. Pressure-treated cells were characterized in order to determine viability, the transmembrane difference in pH ({Delta}pH), and multiple-drug-resistance (MDR) transport activity. Furthermore, pressure effects on the intracellular pH and the fluidity of the membrane were determined during pressure treatment. In the presence of external sucrose and NaCl, high intracellular levels of sucrose and lactose, respectively, were accumulated by L. lactis; 4 M NaCl and, to a lesser extent, 0.5 M sucrose provided protection against pressure-induced cell death. The transmembrane {Delta}pH was reversibly dissipated during pressure treatment in any buffer system. Sucrose but not NaCl prevented the irreversible inactivation of enzymes involved in pH homeostasis and MDR transport activity. In the presence 0.5 M sucrose or 4 M NaCl, the fluidity of the cytoplasmic membrane was maintained even at low temperatures and high pressure. These results indicate that disaccharides protect microorganisms against pressure-induced inactivation of vital cellular components. The protective effect of ionic solutes relies on the intracellular accumulation of compatible solutes as a response to the osmotic stress. Thus, ionic solutes provide only asymmetric protection, and baroprotection with ionic solutes requires higher concentrations of the osmolytes than of disaccharides.

* Corresponding author. Mailing address: TU München, Lehrstuhl Technische Mikrobiologie, Weihenstephaner Steig 16, D-85350 Freising, Germany. Phone: 49 8161 713204. Fax: 49 8161 713327. E-mail: michael.gaenzle{at}wzw.tum.de.

Applied and Environmental Microbiology, April 2004, p. 2013-2020, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2013-2020.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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